The invention relates to the technical field of biological
medicine and vaccine preparation, and discloses a preparation method of a tetravalent influenza
virus split vaccine, and the preparation method comprises the following steps: inoculating four influenza viruses into a chick
embryo, culturing, harvesting
allantoic fluid, and concentrating; centrifugally purifying through a
sucrose density zone, and adding a
cracking agent for
cracking; removing the
cracking agent from the cracking solution through
chromatographic column chromatography, and then adding an inactivating agent for inactivating and removing the inactivating agent; according to the method disclosed by the invention, a process
route of firstly splitting purification and then inactivation in combination with dual purification of
centrifugation and
chromatography is adopted, so that the problem of
impurity cross-linking caused by an inactivator in a traditional process is effectively avoided, and residues of
ovalbumin and splitting agents are deeply removed; and the natural conformation of the
virus antigen is retained to the greatest extent. The obtained vaccine is extremely low in
impurity content, has excellent safety and high
immunogenicity, and can effectively expand the
immune protection range and enhance the
population immune barrier.